High resolution "Ultra performance" liquid chromatography coupled to oa-TOF mass spectrometry as a tool for differential metabolic pathway profiling in functional genomic studies
High resolution "Ultra performance" liquid chromatography coupled to oa-TOF mass spectrometry as a tool for differential metabolic pathway profiling in functional genomic studies
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DOI：
10.1021/pr049769r
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发表时间：
2005-03-01
影响因子：
4.4
通讯作者：
Plumb, RS
中科院分区：
文献类型：
--
作者：
Wilson, ID;Nicholson, JK;Plumb, RS
本文介绍了一种新型的1.7 μ m反相填料与色谱系统的组合。12 000 psi（所谓的超高效液相色谱，UPLC）已经能够显著提高复杂混合物分离的色谱性能。性能的提高表现在峰分辨率的提高以及速度和灵敏度的提高。在这里，我们表明，超高效液相色谱提供了显着的优势，超过传统的反相HPLC相当于一个超过一倍的峰值容量，速度增加近10倍，灵敏度增加3至5倍，与传统的3.5 μ m固定相相比。UPLC-MS技术的第一个功能性基因组应用在这里示出了关于来自两组表型正常小鼠品系（C57 BL 19 J和Alpk：ApfCD）和“裸鼠”品系的雄性和雌性的尿液的多变量代谢谱。我们还在类似的分析条件下将该技术与常规HPLC-MS进行了比较，并显示出UPLC-MS分析的表型分类能力得到改善，同时由于分析灵敏度和分辨率得到改善，探测菌株之间差异途径活性的能力也有所提高。
The combination of a new 1.7 mu m reversed-phase packing material, and a chromatographic system, operating at ca. 12 000 psi, (so-called ultra performance liquid chromatography, UPLC) has enabled dramatic increases in chromatographic performance to be obtained for complex mixture separation. This increase in performance is manifested in improved peak resolution, together with increased speed and sensitivity. Here, we show that UPLC offers significant advantages over conventional reversed-phase HPLC amounting to a more than doubling of peak capacity, an almost 10-fold increase in speed and a 3- to 5-fold increase in sensitivity compared to that generated with a conventional 3.5 mu m stationary phase. The first functional genomic application of UPLC-MS technology is illustrated here with respect to multivariate metabolic profiling of urines from males and females of two groups of phenotypically normal mouse strains (C57BL19J and Alpk:ApfCD) and a "nude mouse" strain. We have also compared this technology to conventional HPLC-MS under similar analytical conditions and show improved phenotypic classification capability of UPLC-MS analysis together with increased ability to probe differential pathway activities between strains as a result of improved analytical sensitivity and resolution.